Desulfoglaeba Medium (DSMZ Medium 1074)

[所属分类:培养基配方] [发布时间:2020-6-5] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Desulfoglaeba Medium
(DSMZ Medium 1074)

培养基配方
Composition per liter:
NaCl .......................................................................20.0g
Na2SO4 ......................................................................3.5g
MgCl2 ·6H2O ................................................................3.0g
KCl .........................................................................0.5g
NH4Cl ......................................................................0.25g
KH2PO4 ......................................................................0.2g
CaCl2 ·2H2O.................................................................0.15g
Yeast extract ...............................................................0.05g
Resazurin ...................................................................0.5mg
Wolfe’s vitamin solution...................................................10.0mL
NaHCO3  solution ...........................................................10.0mL
Na2S·9H2O solution.........................................................10.0mL
L -Cysteine·HCl solution...................................................10.0mL
Wolfe's Mineral Elixir.......................................................1.0mL
n-Decane ....................................................................1.0mL
pH 7.1 ± 0.2 at 25°C
L -Cysteine·HCl Solution:
Composition per 10.0mL:
L -Cysteine·HCl..............................................................0.1g
Preparation of  L -Cysteine Solution : Add  L -cysteine·HCl to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave under 100% N 2 for 15 min at 15 psi
pressure–121°C.
Na2S·9H2O Solution :
Composition per 10.0mL:
Na2S·9H2O....................................................................0.1g
Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
NaHCO3 Solution :
Composition per 10.0mL:
NaHCO3 .....................................................................3.0g
Preparation of NaHCO3 Solution:  Add components to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO2 + 80% H2 . Autoclave for 15 min at 15 psi pressure–
121°C. Cool to room temperature.
Wolfe’s Mineral Elixir:
Composition per liter :
MgSO4 ·7H2O................................................................30.0g
NaCl........................................................................10.0g
MnSO4 ·2H2O.................................................................5.0g
(NH4)2 NiSO4 ·6H2O .........................................................2.8g
CoCl2 ·6H2O ................................................................1.8g
ZnSO4 ·7H2O.................................................................1.8g
FeSO4 ·7H2O.................................................................1.0g
CaCl2 ·2H2O.................................................................1.0g
KAl(SO4)2 ·12H2O...........................................................0.18g
CuSO4 ·5H2O................................................................0.1g
H3BO3 ......................................................................0.1g
Na2MoO4 ·2H2O .............................................................0.1g
Na2SeO4 ....................................................................0.1g
Na2WO4 ·2H2O...............................................................0.1g
Preparation of Wolfe’s Mineral Elixir: Adjust pH of 1.0L of dis-
tilled/deionized water to 1.0 with dilute H2SO4 . Add remaining com-
ponents one at a time. Mix thoroughly to dissolve.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl.........................................................10.0mg
Thiamine·HCl ...........................................................5.0mg
Riboflavin ..............................................................5.0mg
Nicotinic acid...........................................................5.0mg
Calcium pantothenate ....................................................5.0mg
p-Aminobenzoic acid......................................................5.0mg
Thioctic acid............................................................5.0mg
Biotin...................................................................2.0mg
Folic acid ..............................................................2.0mg
Cyanocobalamin........................................................100.0μg
Preparation of Wolfe’s Vitamin Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize.
Preparation of Medium:  Add components, except bicarbonate, vi-
tamins, and reducing agents, to distilled/deionized water and bring vol-
ume to 960.0mL. Adjust pH to 7.2. Gently heat and bring to boiling.
Boil for 1 min. Mix thoroughly. Cool to room temperature while sparg-
ing with 80% N2  + 20% CO2 . Dispense into culture vessels under an
atmosphere of 80% N2  + 20% CO2  (e.g., Balch tubes or serum vials).
Add n-decane to each vessel. Close the vessels using butyl rubber stop-
pers fixed with aluminum crimps. Autoclave for 15 min at 15 psi pres-
sure–121°C. Add vitamins and reducing agents from sterile, anoxic
stock solutions. Adjust pH to 7.1. Use 20% as inoculum and incubate
vials in an inverted position.
Use: For the cultivation of Desulfoglaeba spp.